Novel function of the human presqualene diphosphate phosphatase as a type II phosphatidate phosphatase in phosphatidylcholine and triacylglyceride biosynthesis pathways.
Phosphatidate phosphatases, PAPs, are key enzymes in lipid biosynthesis and signaling. Type I PAP enzymes participate in de-novo phospholipid biosynthesis, whereas type II PAP enzymes have an established role in lipid signaling. To identify novel human type II PAPs potentially involved in de-novo phospholipid synthesis we used bioinformatics to screen for enzymes with an active site exposed to the cytosolic side of membranes. Two related enzymes, a novel lipid phosphatase related protein (LPRP-A) and a presqualene diphosphate phosphatase (PA-PSP) met this criterion. PA-PSP and LPRP-A have differential tissue and subcellular distribution, and novel yet differential roles in lipid metabolism. Specifically, PA-PSP, but not LPRP-A, was a potent Mg(2+)-independent, NEM-insensitive type II PAP. Subcellular fractionation detection indicated that both proteins were associated with membranes, while immunofluorescent deconvolution imaging revealed that these membranes were exclusively from the nuclear envelope and the endoplasmic reticulum. PA-PSP overexpression, but not LPRP-A, accelerated the synthesis of phosphatidylcholine and caused accumulation of triacylglycerol with concomitant decrease in the rate of phosphatidylinositol synthesis. Coexpression of human CTP:phosphocholine cytidylyltransferase-alpha with PA-PSP enhanced the effect of PA-PSP on phosphatidylcholine levels, yet attenuated its effect on triacylglycerol. Taken together, our studies provide the first evidence that the eukaryotic, ER-resident PA-PSP is a bifunctional enzyme with specific type II PAP activity, and regulates, in addition to type I PAPs, the de-novo biosynthesis of phospholipids and triacylglycerols.
Theofilopoulos
Spyros
S
Center for Neurosciences, Biomedical Research Foundation of the Academy of Athens, Athens 11527, Greece.
Lykidis
Athanasios
A
Leondaritis
George
G
Mangoura
Dimitra
D
D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D002784:Cholesterol / Q000378:metabolism; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010711:Phosphatidate Phosphatase / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012394:Rosaniline Dyes / Q000737:chemistry; D013347:Subcellular Fractions; D014280:Triglycerides / Q000096:biosynthesis*
D009419:Nerve Tissue Proteins; D010713:Phosphatidylcholines; D011106:Polyisoprenyl Phosphates; D012333:RNA, Messenger; D012394:Rosaniline Dyes; D014280:Triglycerides; C005095:malachite green; C017672:presqualene pyrophosphate; D002784:Cholesterol; C486236:PLPPR2 protein, human; C406242:phosphatidic acid phosphatase type 2; D010744:Phosphoric Monoester Hydrolases; D010711:Phosphatidate Phosphatase